Cross-species single-cell RNA sequencing reveals divergent phenotypes and activation states of adaptive immunity in human carotid and experimental murine atherosclerosis

  • Cardiovasc Res. 2024 Nov 25;120(14):1713-1726. doi: 10.1093/cvr/cvae154.
Hauke Horstmann  1  2  3 ,  Nathaly Anto Michel  4 ,  Xia Sheng  1  2 ,  Sophie Hansen  1  2 ,  Alexandra Lindau  1  2 ,  Katharina Pfeil  3 ,  Marbely C Fernández  2  5 ,  Timoteo Marchini  1  2 ,  Holger Winkels  6 ,  Lucia Sol Mitre  1  2  7 ,  Tijani Abogunloko  1  2  7 ,  Xiaowei Li  1  2 ,  Timothy Bon-Nawul Mwinyella  1  2 ,  Mark Colin Gissler  1  2 ,  Heiko Bugger  2  4 ,  Timo Heidt  1  2 ,  Konrad Buscher  8 ,  Ingo Hilgendorf  1  2 ,  Peter Stachon  1  2 ,  Sven Piepenburg  1  2 ,  Nicolas Verheyen  4 ,  Thomas Rathner  4 ,  Teresa Gerhardt  9  10  11 ,  Patrick Malcolm Siegel  1  2 ,  Wolfgang Kurt Oswald  12 ,  Tina Cohnert  12 ,  Alma Zernecke  13 ,  Josef Madl  2  5 ,  Peter Kohl  2  5 ,  Amanda C Foks  14 ,  Constantin von Zur Muehlen  1  2 ,  Dirk Westermann  1  2 ,  Andreas Zirlik  4 ,  Dennis Wolf  1  2
Affiliations
  • 1. Department of Cardiology and Angiology I, Medical Center, University of Freiburg, 79106 Freiburg, Germany.
  • 2. Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany.
  • 3. Division of Cardiology, Department of Medicine, NYU Cardiovascular Research Center, NYU Grossmann School of Medicine, 10016 New York, NY, USA.
  • 4. Department of Cardiology, University Heart Center Graz, Medical University of Graz, 8036 Graz, Austria.
  • 5. Institute for Experimental Cardiovascular Medicine, Heart Centre, University of Freiburg, 79106 Freiburg, Germany.
  • 6. Department of Cardiology, Faculty of Medicine and University Hospital Cologne, University of Cologne, 50923 Cologne, Germany.
  • 7. Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, 79106 Freiburg, Germany.
  • 8. Division of General Internal Medicine, Nephrology and Rheumatology, Department of Medicine, University Hospital of Münster, 48149 Münster, Germany.
  • 9. Department of Cardiology, Angiology and Intensive Care Medicine CBF, Deutsches Herzzentrum der Charité, and Berlin Institute of Health (BIH), 13353 Berlin, Germany.
  • 10. DZHK (German Centre for Cardiovascular Research), partner site Berlin , 13353 Berlin, Germany.
  • 11. Cardiovascular Research Institute and the Department of Medicine, Cardiology, Icahn School of Medicine at Mount Sinai, 10029 New York, NY, USA.
  • 12. Department of Vascular Surgery, Medical University of Graz, 8036 Graz, Austria.
  • 13. Institute of Experimental Biomedicine, University Hospital Würzburg, 97080 Würzburg, Germany.
  • 14. Leiden Academic Centre for Drug Research, Division of BioTherapeutics, Leiden University, 2333 CC Leiden, The Netherlands.
PMID: 39041203 DOI: 10.1093/cvr/cvae154
Abstract

Aims: The distinct functions of immune cells in atherosclerosis have been mostly defined by pre-clinical mouse studies. Contrastingly, the immune cell composition of human atherosclerotic plaques and their contribution to disease progression are only poorly understood. It remains uncertain whether genetic animal models allow for valuable translational approaches.

Methods and results: Single-cell RNA-sequencing (scRNA-seq) was performed to define the immune cell landscape in human carotid atherosclerotic plaques. The human immune cell repertoire demonstrated an unexpectedly high heterogeneity and was dominated by cells of the T-cell lineage, a finding confirmed by immunohistochemistry. Bioinformatical integration with 7 mouse scRNA-seq data sets from adventitial and atherosclerotic vascular tissue revealed a total of 51 identities of cell types and differentiation states, of which some were only poorly conserved between species and exclusively found in humans. Locations, frequencies, and transcriptional programmes of immune cells in mouse models did not resemble the immune cell landscape in human carotid atherosclerosis. In contrast to standard mouse models of atherosclerosis, human plaque leucocytes were dominated by several T-cell phenotypes with transcriptional hallmarks of T-cell activation and memory formation, T-cell receptor, and pro-inflammatory signalling. Only mice at the age of 22 months partially resembled the activated T-cell phenotype. In a validation cohort of 43 patients undergoing carotid endarterectomy, the abundance of activated immune cell subsets in the plaque defined by multi-colour flow cytometry associated with the extent of clinical atherosclerosis.

Conclusion: Integrative scRNA-seq reveals a substantial difference in the immune cell composition of murine and human carotid atherosclerosis-a finding that questions the translational value of standard mouse models for adaptive immune cell studies. Clinical associations suggest a specific role for T-cell driven (auto-)immunity in human plaque formation and instability.

Keywords
Atherosclerosis; Immunity; Leucocytes; Prediction; ScRNA-seq; Transcriptome.